Freeze-drying technique,also known as lyophilization,is a process of removing moisture from a solution or suspension through freezing and vacuum dehydration to maintain the stability of the samples and prolong their shelf life.Freeze-drying technology has been widely used in food,pharmaceutical,clinical testing,chemical and other fields but its application in the field of forensic medicine has just started.Polymerase chain reaction(PCR)and its derivative detection technologies are widely used in forensic DNA detection,but PCR reagents need to be stored and transported at low temperature.In recent years,forensic scientists have begun applying freeze-drying technology to PCR amplification reagents to solve the transportation and storage problems of PCR reagents.In order to promote the application of PCR freeze-drying technology in forensic genetics,this paper mainly expounds the research course,system and process of PCR freeze-drying technology,compares the advantages and disadvantages of PCR reagents with traditional PCR reagents,and introduces the advantages and challenges of PCR freeze-drying reagents in forensic medicine.

Freeze-drying technique,also known as lyophilization,is a process of removing moisture from a solution or suspension through freezing and vacuum dehydration to maintain the stability of the samples and prolong their shelf life.Freeze-drying technology has been widely used in food,pharmaceutical,clinical testing,chemical and other fields but its application in the field of forensic medicine has just started.Polymerase chain reaction(PCR)and its derivative detection technologies are widely used in forensic DNA detection,but PCR reagents need to be stored and transported at low temperature.In recent years,forensic scientists have begun applying freeze-drying technology to PCR amplification reagents to solve the transportation and storage problems of PCR reagents.In order to promote the application of PCR freeze-drying technology in forensic genetics,this paper mainly expounds the research course,system and process of PCR freeze-drying technology,compares the advantages and disadvantages of PCR reagents with traditional PCR reagents,and introduces the advantages and challenges of PCR freeze-drying reagents in forensic medicine.

To explore the mechanism of Suanzaoren Decoction in the treatment of insomnia from endogenous bile acid regulation, the present study investigated the hepatoprotective effect of Suanzaoren Decoction and the molecular changes of bile acids in the serum, liver, and ileum of insomnia model mice and Suanzaoren Decoction treated mice. The insomnia model in mice was established by the sleep deprivation method. After Suanzaoren Decoction(48.96 mg·kg~(-1)·d~(-1)) intervention by gavage for 7 days, the related indicators, such as water consumption, food intake, body weight, aspartate aminotransferase(AST), alanine transaminase(ALT), and total bile acid(TBA) were detected, and the pathological changes of the liver and ileum were observed. The molecular levels and distribution of 23 bile acids in the serum, liver, and ileum were analyzed by UPLC-MS/MS combined with principal component analysis(PCA) and partial least squares discriminant analysis(PLS-DA). The results showed that Suanzaoren Decoction could improve the decreased water consumption and food intake, weight loss, and increased AST and ALT in the model group, and effectively reverse the injury and inflammation in the liver and ileum. The bile acids in the liver of the insomnia model mice were in the stage of decompensation, and the bile acids in the serum, liver, and ileum of the mice decreased or increased. Suanzaoren Decoction could regulate the anomaly of some bile acids back to normal. Seven bile acids including glycoursodeoxycholic acid(GUDCA), glycodesoxycholic acid(GDCA), tauro-α-MCA(T-α-MCA), α-MCA, taurodeoxycholate(TDCA), T-β-MCA, and LCA were screened out as the main discriminant components by PLS-DA. It is concluded that Suanzaoren Decoction possesses the hepatoprotective effect and bile acids could serve as the biochemical indicators to evaluate the drug efficacy in the treatment of abnormal liver functions caused by insomnia. The mechanism of Suanzao-ren Decoction in soothing the liver, resolving depression, tranquilizing the mind, and improving sleep may be related to the molecular regulation of bile acid signals.
Animals ; Bile Acids and Salts ; Chromatography, Liquid ; Drugs, Chinese Herbal ; Ileum ; Liver ; Mice ; Sleep Initiation and Maintenance Disorders/drug therapy* ; Tandem Mass Spectrometry

OBJECTIVES: To investigate the clinical treatment outcomes and the changes of the outcomes over time in extremely preterm twins in Guangdong Province, China. METHODS: A retrospective analysis was performed for 269 pairs of extremely preterm twins with a gestational age of <28 weeks who were admitted to the department of neonatology in 26 grade A tertiary hospitals in Guangdong Province from January 2008 to December 2017. According to the admission time, they were divided into two groups: 2008-2012 and 2013-2017. Besides, each pair of twins was divided into the heavier infant and the lighter infant subgroups according to birth weight. The perinatal data of mothers and hospitalization data of neonates were collected. The survival rate of twins and the incidence rate of complications were compared between the 2008-2012 and 2013-2017 groups. RESULTS: Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of severe asphyxia and smaller head circumference at birth (P<0.05). The mortality rates of both of the twins, the heavier infant of the twins, and the lighter infant of the twins were lower in the 2013-2017 group compared with the 2008-2012 group (P<0.05). Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of pulmonary hemorrhage, patent ductus arteriosus (PDA), periventricular-intraventricular hemorrhage (P-IVH), and neonatal respiratory distress syndrome (NRDS) and a higher incidence rate of bronchopulmonary dysplasia (P<0.05). CONCLUSIONS There is a significant increase in the survival rate over time in extremely preterm twins with a gestational age of <28 weeks in the 26 grade A tertiary hospitals in Guangdong Province. The incidences of severe asphyxia, pulmonary hemorrhage, PDA, P-IVH, and NRDS decrease in both the heavier and lighter infants of the twins, but the incidence of bronchopulmonary dysplasia increases. With the improvement of diagnosis and treatment, the multidisciplinary collaboration between different fields of fetal medicine including prenatal diagnosis, obstetrics, and neonatology is needed in the future to jointly develop management strategies for twin pregnancy.
Bronchopulmonary Dysplasia/epidemiology* ; Female ; Gestational Age ; Humans ; Infant ; Infant, Extremely Premature ; Infant, Newborn ; Pregnancy ; Respiratory Distress Syndrome, Newborn/epidemiology* ; Retrospective Studies ; Treatment Outcome

Reduced levels of retinal dopamine, a key regulator of eye development, are associated with experimental myopia in various species, but are not seen in the myopic eyes of C57BL/6 mice, which are deficient in melatonin, a neurohormone having extensive interactions with dopamine. Here, we examined the relationship between form-deprivation myopia (FDM) and retinal dopamine levels in melatonin-proficient CBA/CaJ mice. We found that these mice exhibited a myopic refractive shift in form-deprived eyes, which was accompanied by altered retinal dopamine levels. When melatonin receptors were pharmacologically blocked, FDM could still be induced, but its magnitude was reduced, and retinal dopamine levels were no longer altered in FDM animals, indicating that melatonin-related changes in retinal dopamine levels contribute to FDM. Thus, FDM is mediated by both dopamine level-independent and melatonin-related dopamine level-dependent mechanisms in CBA/CaJ mice. The previously reported unaltered retinal dopamine levels in myopic C57BL/6 mice may be attributed to melatonin deficiency.
Animals ; Disease Models, Animal ; Dopamine ; Melatonin ; Mice ; Mice, Inbred C57BL ; Mice, Inbred CBA ; Myopia ; Retina ; Sensory Deprivation

To investigate the effects of anti-androgen drugs and melengestrol acetate (MGA) on development of regrowth antlers in 6 year old sika deer, twenty healthysika deerwith similar body weight and antler weightwere randomly divided into five groups by using single factor test design: flutamide (=4), bicalutamide (=4), progesterone acetate (CPA, =4), melengestrol acetate (MGA, =4), control(=4). All deer were fed with same diets and were housed outside together in an opened fence of 15 m×30 m with free access to water and feed. Treatment groups were injected subcutaneously sustained-release agents of the four drugs respectively when two-branched antlers were harvested. The control group had no special treatment. In the experiment period of 60 d, blood sampleswere collected for 4 times for each deer. The concentration of testosterone in plasma was tested and analyzed to compare the changes between different groups. Development of regrowth antlers was observed. At the end of the experiment, regrowth antlers were weighted and analyzed. The resultsshowed that the weights of regrowth antlers in treatment groups were significantly greater than those from control group and the weight gain (as compared with the control group) was 100.50%, 64.46%, 87.16% and 117.46% respectively in flutamide group, bicalutamide group, progesterone acetate group and melengestrol acetate group. For plasma testosterone concentration, it was not significantly different in the early stage (in the first 35 d), but at the end of the experimen, it was significantly higher than that of earlier stage (<0.01) in various groups. Testosterone concentration of flutamide treated group was significantly lower than that of the other groups (<0.01), while the level inbicalutamide and MGA treated groups was significantly higher than that in other groups (<0.01). The results showed that both anti-androgen drugs and MGA treatment promoted the development of regrowth antlers and increased the weight of regrowth antlers, where the effect was most significant by MGA treatment. From the morphological observation of the antlers, it was found that anti-androgen and MGA treatments prolonged the growth period of regrowth antlers through delaying the ossification of antlers. However, plasma testosterone concentration was not affected by the treatments.

OBJECTIVETo compare technique and clinical effect of arthroscopic and conventional sectional therapy for the treatment of ischial tuberosity cyst.

METHODSFrom May 2014 and September 2016, 49 patients with symptomatic ischial tuberosity cyst were randomized divided into arthroscopic group and conventional section group by envelope method. There were 24 patients in arthroscopic group, including 16 males and 8 females aged from 42 to 81years old with an average age of (64.1±9.3) years old;the courses of disease ranged from 2 to 36 months with an average of (17.0±9.1) months;treated with removing cyst wall under arthroscopic and build artificial lacuna around cyst. There were 25 patients in conventional section group, including 11 males and 14 females aged from 47 to 79 years old with an average of (61.2±10.6) years old; the courses of disease ranged from 4 to 36 months with an average of (17.5±8.5) months;treated with cutting off lump with transverse incision. Operative time, blood loss, fluid volume, hospital stays and postoperative complication were observed and compared, VAS score were applied to evaluate pain degree.

RESULTSForty-nine patients were followed up from 6 to 18 months with an average of (11.3±3.3) months. In conventional group, 2 patients occurred incision infection, 1 case reoccurred. All patients in arthroscopic group were healed at stage I. Operative time, blood loss, fluid volume, and hospital stays in arthroscopic group were (54.7±7.7) min, (20.8±3.5) ml, (20.3±5.6) ml, (2.8±0.6) d respectively and better than that of conventional group(71.8±8.8) min, (67.3±12.0) ml, (103.6±20.3) ml, (7.8±2.9) d. Postoperative VAS score in arthroscopic group on the first day, seventh days and first month were 2.6±0.7, 0.5±0.6, 0.3±0.5, and significantly lower than that of conventional group 6.0±0.7, 3.0±1.0, 1.1±1.0 respectively(<0.05). There were no statistical difference in postoperative complications between two groups(>0.05).

CONCLUSIONSCompared with conventional group, arthroscopic for ischial tuberosity cyst has advantages of minimal invasive, less blood loss during perioperative period, less pain degree after surgery, safety and rapid recovery. But arthroscopic skills are needed for surgeons. It should be recommended in clinical practice.

Adult ; Aged ; Aged, 80 and over ; Arthroscopy ; Bone Cysts ; surgery ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Operative Time ; Pelvic Bones ; pathology ; Postoperative Period ; Prospective Studies ; Treatment Outcome

Aim To investigate the effect of broneol on acute lung injury(ALI) induced by lipopolysaccharide (LPS). Methods Male C57 mice were randomly di-vided into saline group, model group, broneol group and dexamethasone group, then the ALI mouse model was induced by instilling intratracheally with LPS. The levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6) and keratinocyte-de-rived cytokine (KC) were measured at 6h, 12h and 24h after instillation of LPS, and the pathological changes of lung were observed. Mice alveolar macro-phages (MHS) and epithelial cells (MLE-12) were stimulated by LPS. After the stimulation of 1h, 3h, 6h,9h, 12h, 24h, the levels of TNF-α and IL-6 in MHS cells and the contents of KC and macrophage in-flammatory protein-2 (MIP-2) in MLE-12 cells were measured. Results Broneol could inhibit the secre-tion of TNF-α,KC and IL-1β;the early effect of bro-neol on IL-6 was not obvious,but the later effect after the treatment of 24 hours was obvious. After LPS instil-lation 6h and 12h,Broneol could significantly improve lung tissue pathological changes. Broneol had no effect on TNF-α secretion of MHS cells, but it obviously af-fected IL-6 secretion in the later stage. In addition, broneol significantly inhibited KC and MIP2 secretion in MLE-12 cells at the later stage of LPS stimulation. Conclusions Broneol can protect LPS-induced acute lung injury. The mechanism may be related to the inhi-bition of the release of inflammatory factors,the activa-tion of inflammatory cells and the aggregation of neutro-phils.

BACKGROUNDCirculating endometrial cells (CECs) have been reported to be present in the peripheral blood of women with endometriosis (EM), providing clear and specific evidence of the presence of ectopic lesions. In this study, we established a method with a high detection rate of CECs, assessed the diagnostic value of CECs for EM and compared with serum CA125, and proposed a hypothesis for the pathogenesis of EM from the new perspective of CECs.

METHODSThe participants were enrolled prospectively from October 2015 to July 2016. The peripheral blood samples were collected from 59 participants, and the blood cells were isolated for immunofluorescence staining via microfluidic chips. The cells that were positive for vimentin/cytokeratin and estrogen/progesterone receptor and negative for CD45 were identified as CECs. The serum CA125 level was tested with electrochemiluminescence immunoassay.

RESULTSThe detection rate of CECs reached 89.5% (17/19) in the EM group, which was significantly higher than that of the control group (15.0% [6/40], P < 0.001) and was independent of menstrual cycle phases. Furthermore, a positive CEC assay detected 4/5 cases of Stage I-II EM. In contrast, a positive CA125 test had limited value in detecting EM (13/19, 68.4%) and detected only one case of Stage I-II EM.

CONCLUSIONCECs are promising biomarkers for EM with great potential for a noninvasive diagnostic assay.

AIM To prepare diosgenin nanosuspensions.METHODS The nanosuspensions prepared by media milling method were solidificated by freeze drying method.With particle size and polydispersity index (PDI) as evaluation indices,stabilizer kind,ratio of diosgenin to stabilizer,ratio of preliminary nanosuspension volume to grinding bead amount,milling time,lyoprotectant kind and its amount as influencing factors,single factor test was applied to screening preparation and solidification processes.The morphology of nanosuspensions was observed,then the particle sizes and polydispersity indices of nanosuspensions and freeze-dried powder were determined.RESULTS The optimal conditions were determined to be 6:1 for ratio of diosgenin to Pluronic F127 (stabilizer Ⅰ),50:1 for ratio of diogenin to sodium dodecyl sulfate (SDS,stabilizer Ⅱ),1:4 for ratio of preliminary nanosuspension volume to grinding bead amount,120 min for milling time,and 8％ PEG-6000 and 2％ mannitol as lyoprotectants.The average particle size and polydispersity index of rod-like or flaky nanosuspensions were (348.1 ±14.2) nm and 0.244 ± 0.059,respectively,which were lower than those of freeze-dried powder.At room temperature,the particle sizes of nanosuspensions and freeze-dried powder remained stable within 35 d and 3 months,respectively.CONCLUSION The physical stability of diosgenin freeze-dried powder is superior to that of its nanosuspensions,which can be used after being reconstituted.